Specific Serum Antibodies Research Articles

Severe acute respiratory syndrome coronavirus 2 prevalence in saliva and gastric and intestinal fluid in patients undergoing gastrointestinal endoscopy in coronavirus disease 2019 endemic areas: Prospective cross-sectional study in Japan.

ObjectivesGastrointestinal endoscopy (GIE) is useful for the early detection and treatment of many diseases; however, GIE is considered a high‐risk procedure in the coronavirus disease 2019 (COVID‐19) pandemic era. This study aimed to explore the rate of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) positivity in saliva and gastrointestinal fluids to which endoscopy medical staff are exposed.MethodsThe study was a single‐center cross‐sectional study. From June 1 to July 31, 2020, all patients who underwent GIE at Yokohama City University Hospital were registered. All patients provided 3 mL of saliva. For upper GIE, 10 mL of gastric fluid was collected through the endoscope. For lower GIE, 10 mL of intestinal fluid was collected through the endoscope. The primary outcome was the positive rate of SARS‐CoV‐2 in saliva and gastrointestinal fluids. We also analyzed serum‐specific antibodies for SARS‐CoV‐2 and patients’ background information.ResultsA total of 783 samples (560 upper GIE and 223 lower GIE samples) were analyzed. Polymerase chain reaction (PCR) on saliva samples did not show any positive results in either upper or lower GIE samples. However, 2.0% (16/783) of gastrointestinal fluid samples tested positive for SARS‐CoV‐2. No significant differences in age, sex, purpose of endoscopy, medication, or rate of antibody test positivity were found between PCR positive and PCR negative cases.ConclusionsAsymptomatic patients, even those with no detectable virus in their saliva, had SARS‐CoV‐2 in their gastrointestinal tract. Endoscopy medical staff should be aware of infection when performing procedures. The study was registered as UMIN000040587.

Bovine Holo-Beta-Lactoglobulin Cross-Protects Against Pollen Allergies in an Innate Manner in BALB/c Mice: Potential Model for the Farm Effect.

The lipocalin beta-lactoglobulin (BLG) is a major protein compound in cow’s milk, and we detected it in cattle stable dust. BLG may be a novel player in the farm protective effect against atopic sensitization and hayfever. In previous studies, we demonstrated that only the ligand-filled holo-form of BLG prevented sensitization to itself. Here, we investigated whether holo-BLG could, in an innate manner, also protect against allergic sensitization to unrelated birch pollen allergens using a murine model. BALB/c mice were nasally pretreated four times in biweekly intervals with holo-BLG containing quercetin–iron complexes as ligands, with empty apo-BLG, or were sham-treated. Subsequently, mice were intraperitoneally sensitized two times with apo-BLG or with the unrelated birch pollen allergen apo-Bet v 1, adjuvanted with aluminum hydroxide. After subsequent systemic challenge with BLG or Bet v 1, body temperature drop was monitored by anaphylaxis imaging. Specific antibodies in serum and cytokines of BLG- and Bet v 1-stimulated splenocytes were analyzed by ELISA. Additionally, human peripheral blood mononuclear cells of pollen allergic subjects were stimulated with apo- versus holo-BLG before assessment by FACS. Prophylactic treatment with the holo-BLG resulted in protection against allergic sensitization and clinical reactivity also to Bet v 1 in an unspecific manner. Pretreatment with holo-BLG resulted in significantly lower BLG-as well as Bet v 1-specific antibodies and impaired antigen-presentation with significantly lower numbers of CD11c+MHCII+ cells expressing CD86. Pretreatment with holo-BLG also reduced the release of Th2-associated cytokines from Splenocytes in BLG-sensitized mice. Similarly, in vitro stimulation of PBMCs from birch pollen allergic subjects with holo-BLG resulted in a relative decrease of CD3+CD4+ and CD4+CRTh2 cells, but not of CD4+CD25+CD127− Treg cells, compared to apo-BLG stimulation. In conclusion, prophylactic treatment with holo-BLG protected against allergy in an antigen-specific and -unspecific manner by decreasing antigen presentation, specific antibody production and abrogating a Th2-response. Holo-BLG therefore promotes immune resilience against pollen allergens in an innate manner and may thereby contribute to the farm protective effect against atopic sensitization.

Virus-specific memory T cell responses unmasked by immune checkpoint blockade cause hepatitis

Treatment of advanced melanoma with combined PD-1/CTLA-4 blockade commonly causes serious immune-mediated complications. Here, we identify a subset of patients predisposed to immune checkpoint blockade-related hepatitis who are distinguished by chronic expansion of effector memory CD4+ T cells (TEM cells). Pre-therapy CD4+ TEM cell expansion occurs primarily during autumn or winter in patients with metastatic disease and high cytomegalovirus (CMV)-specific serum antibody titres. These clinical features implicate metastasis-dependent, compartmentalised CMV reactivation as the cause of CD4+ TEM expansion. Pre-therapy CD4+ TEM expansion predicts hepatitis in CMV-seropositive patients, opening possibilities for avoidance or prevention. 3 of 4 patients with pre-treatment CD4+ TEM expansion who received αPD-1 monotherapy instead of αPD-1/αCTLA-4 therapy remained hepatitis-free. 4 of 4 patients with baseline CD4+ TEM expansion given prophylactic valganciclovir and αPD-1/αCTLA-4 therapy remained hepatitis-free. Our findings exemplify how pathogen exposure can shape clinical reactions after cancer therapy and how this insight leads to therapeutic innovations.

L’herpès génital

AbstractGenital herpes is the main cause of genital ulcers worldwide. It is caused by highly prevalent viruses in the general population: herpes simplex viruses 1 and 2 (HSV-1 and HSV-2). This infection may affect quality of life of infected individuals. Genital herpes is a cofactor of sexual transmission of human immunodeficiency virus (HIV). Vertical transmission of HSV may result in neonatal herpes. Biological diagnosis of genital herpes is performed by the detection of viral genome by PCR in genital swabs and the detection of specific antibodies in sera (serology). Therapeutic management of genital herpes is based mainly on antiviral treatments by valaciclovir, very powerful molecule with almost no side effect. To date, there is no vaccine and the use of condoms remains the best way to prevent this viral infection.

Tocilizumab for Severe COVID-19 Pneumonia: Experience from 5 Geriatric Chinese Patients with 6 Months Follow-up.

To enable physicians to understand the efficacy and safety of Tocilizumab (TCZ) in patients with severe coronavirus disease-2019 (COVID-19). We respectively reviewed the clinical records, laboratory results, and chest computed tomography (CT) scans of 5 geriatric patients with severe COVID-19 treated with TCZ during their inpatient hospitalization period in Wuhan from February 08, 2020 to April 04, 2020. The survival status of the patients in the third and the sixth month after being discharged was followed up and recorded. On the fourteenth day after TCZ administration, periphery oxygen saturation rate (SpO2) returned to normal in 4 patients. The serum Interleukin-6 (IL-6) levels altered in five patients after TCZ infusion. One patient rapidly progressed to acute respiratory distress syndrome (ARDS) and died of multiple organ failures eventually. The other 4 patients were cured and discharged from the hospital. During the inpatient hospitalization period, two patients suffered from virus shedding periods (VSPs) delay, and one patient had mild upper respiratory tract infection. One patient died of esophageal carcinoma one month after being discharged. The other 3 patients survived despite mild cough and insomnia. Serum-specific IgG type antibody titer was decreased in one patient. Six months after being discharged, the other three patients were in good condition. TCZ may be an efficient therapeutic option for patients with COVID-19. However, the possibility of VSPs delay, secondary infection, serum protective antibody tilter attenuation, and long-term survival status should be addressed before TCZ therapy initiation.

Persistence of Antibodies to Severe Acute Respiratory Syndrome Coronavirus 2 in Relation to Symptoms in a Nationwide Prospective Study.

BackgroundAssessing the duration of immunity following infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a first priority to gauge the degree of protection following infection. Such knowledge is lacking, especially in the general population. Here, we studied changes in immunoglobulin isotype seropositivity and immunoglobulin G (IgG) binding strength of SARS-CoV-2–specific serum antibodies up to 7 months following onset of symptoms in a nationwide sample.MethodsParticipants from a prospective representative serological study in the Netherlands were included based on IgG seroconversion to the spike S1 protein of SARS-CoV-2 (N = 353), with up to 3 consecutive serum samples per seroconverted participant (N = 738). Immunoglobulin M (IgM), immunoglobulin A (IgA), and IgG antibody concentrations to S1, and increase in IgG avidity in relation to time since onset of disease symptoms, were determined.ResultsWhile SARS-CoV-2–specific IgM and IgA antibodies declined rapidly after the first month after disease onset, specific IgG was still present in 92% (95% confidence interval [CI], 89%–95%) of the participants after 7 months. The estimated 2-fold decrease of IgG antibodies was 158 days (95% CI, 136–189 days). Concentrations were sustained better in persons reporting significant symptoms compared to asymptomatic persons or those with mild upper respiratory complaints only. Similarly, avidity of IgG antibodies for symptomatic persons showed a steeper increase over time compared with persons with mild or no symptoms (P = .022).ConclusionsSARS-CoV-2–specific IgG antibodies persist and show increasing avidity over time, indicative of underlying immune maturation. These data support development of immune memory against SARS-CoV-2, providing insight into protection of the general unvaccinated part of the population.Clinical Trials RegistrationNL8473 (the Dutch trial registry).

Molecular characterization and serodiagnostic potential of Echinococcus granulosus hexokinase

BackgroundCystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus (sensu stricto), is a life-threatening but neglected zoonosis. Glycolytic enzymes are crucial molecules for the survival and development of E. granulosus. The aim of this study was to investigate the molecular characterization, immunogenicity, tissue distribution and serodiagnostic potential of E. granulosus hexokinase (EgHK), the first key enzyme in the glycolytic pathway.MethodsEgHK was cloned and expressed in Escherichia coli. Specific serum antibodies were evaluated in mice immunized with recombinant EgHK (rEgHK). The location of EgHK in the larval stage of E. granulosus was determined using fluorescence immunohistochemistry, and the potential of rEgHK as a diagnostic antigen was investigated in patients with CE using indirect enzyme-linked immunosorbent assay (ELISA).ResultsRecombinant EgHK could be identified in the sera of patients with CE and in mouse anti-rEgHK sera. High titers of specific immunoglobulin G were induced in mice after immunization with rEgHK. EgHK was mainly located in the tegument, suckers and hooklets of protoscoleces and in the germinal layer and laminated layer of the cyst wall. The sensitivity and specificity of the rEgHK-ELISA reached 91.3% (42/46) and 87.8% (43/49), respectively.ConclusionsWe have characterized the sequence, structure and location of EgHK and investigated the immunoreactivity, immunogenicity and serodiagnostic potential of rEgHK. Our results suggest that EgHK may be a promising candidate for the development of vaccines against E. granulosus and an effective antigen for the diagnosis of human CE.Graphical

Detection and analysis of serum food-specific IgG antibody in Beijing area

Objective: The level of food specific IgG antibody in serum of patients with suspected food intolerance in Beijing area was detected and analyzed, so as to provide a basis for patients to eliminate the food that causes disease in their diet. Methods: Data were retrospectively analyzed in a total of 2368 suspected chronic food intolerance patients (all resident population in Beijing) admitted to Beijing Jishuitan Hospital from October 2015 to December 2019, including 859 males and 1509 females. The age range ranged from 2 months to 90 years, (40.2±18.6) years, and the median age was 36 years. Serum specific IgG (sIgG) antibody concentrations in 14 kinds of food (beef, chicken, pork, cod, shrimp, crab, egg, milk, corn, wheat, mushroom, soybean, tomato and rice) were determined by enzym-linked immunosorbentassay (ELISA).SPSS V22.0 was used for statistical analysis, and χ2 was used to compare the positive rate of each group. Results: The positive rate of food sIgG in 2 368 patients was 82.7% (1959/2368), the higher sIgG positive rates were found in eggs (54.7%), shrimp (29.0%) and milk (26.0%), and the lowest sIgG positive rates were found in pork (4.1%) and beef (1.2%) (P<0.05).There was significant gender difference in egg (χ²=10.885) and shrimp (χ²=6.607) sIgG, and female was significantly higher than male (P<0.05).The sIgG positive rates of egg (χ²=515.645), shrimp (χ²=74.20.4), milk (χ²=305.65), soybean (χ²=39.413), wheat (χ²=133.222), cod (χ²=25.540), crab (χ²=55.521), corn (χ²=20.645), tomato (χ²=118.108), chicken (χ²=32.821), pork (χ²=26.539), beef (χ²=21.632) and rice (χ²=25.335) were significant in different age groups. The positive rate of shrimp and crab sIgG increased with age (P<0.05).The positive rate of tomato sIgG decreased with age, and then increased slightly after 45 years. However, egg, milk, soybean, wheat, corn, and rice all showed a decreasing positive rate with the increase of age. The increase of chicken, pork and cod was observed in childhood, and then decreased with age, and then increased in middle and old age. Conclusions: There may be a certain correlation between food intolerance and elevated food sIgG levels in Beijing area. It is suggested that patients of different genders and ages should be optimized and adjusted according to their own characteristics, and children patients are still the key population for food intolerance control.

Prototype of a DNA Vaccine against Marburg Virus.

The preparation and study of the biological properties of the pVAKS-GPVM DNA immunogen containing a gene encoding Marburgvirus glycoprotein are described. The specificity of blood serum antibodies of guinea pigs immunized with DNA immunogen was analyzed by ELISA. Inactivated viral preparation, recombinant glycoprotein (GP) obtained in the prokaryotic system and virus-like particles based on the recombinant vesicular stomatitis virus exhibiting Marburgvirus GP were used as the antigens. The neutralizing activity of antibodies of immunized animals was tested in vitro using a pseudovirus system. It was demonstrated that the developed immunogen administered to guinea pigs induced the production of specific antibodies that neutralize virus-like particles and Marburgvirus in cultured Vero cells.

A prospective surveillance study on the kinetics of the humoral immune response to the respiratory syncytial virus fusion protein in adults in Houston, Texas

Respiratory syncytial virus (RSV)-specific serum antibody has been correlated to protection of infection and reduction of severe disease, but reinfection is still frequent. In this study, we evaluated RSV-specific serum antibody activity following natural RSV re-infection to examine the longevity of the humoral immune response in adults. Nineteen healthy adult volunteers under sixty-five years of age were enrolled during the 2018–2019 RSV season in Houston, TX. Blood was collected at three study visits. The kinetics of RSV-neutralizing, RSV F site-specific competitive, and RSV-binding antibodies in serum samples were measured by microneutralization and enzyme-linked immunosorbent assays. Three distinct profiles of RSV-specific antibody kinetics were identified that were consistent with RSV infection status: uninfected, acutely infected, and recently infected. The uninfected group had stable antibody titers for the duration of the study period (185 days). The acutely infected group had lower antibody responses at the beginning of the study, supporting a correlate of infection, followed by a significant antibody response after infection that was maintained for at least 125 days. Unlike the acutely infected group, the recently infected group had a significant precipitous decrease in RSV antibody in only 60 days. This study is the first, to our knowledge, to describe this abrupt loss of RSV-specific antibody in detail. This rapid decline of antibody may present an obstacle for the development of vaccines with lasting protection against RSV, and perhaps other respiratory pathogens. Neutralizing antibody responses were greater to prototypic than contemporaneous RSV strains, regardless of infection status, indicating that original antigenic sin may impact the humoral immune response to new or emerging RSV strains.

Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany

School and daycare closures were enforced as measures to confine the novel coronavirus disease 2019 (COVID-19) pandemic, based on the assumption that young children may play a key role in severe acute respiratory coronavirus 2 (SARS-CoV-2) spread. Given the grave consequences of contact restrictions for children, a better understanding of their contribution to the COVID-19 pandemic is of great importance. To describe the rate of SARS-CoV-2 infections and the seroprevalence of SARS-CoV-2 antibodies in children aged 1 to 10 years, compared with a corresponding parent of each child, in a population-based sample. This large-scale, multicenter, cross-sectional investigation (the COVID-19 BaWü study) enrolled children aged 1 to 10 years and a corresponding parent between April 22 and May 15, 2020, in southwest Germany. Potential exposure to SARS-CoV-2. The main outcomes were infection and seroprevalence of SARS-CoV-2. Participants were tested for SARS-CoV-2 RNA from nasopharyngeal swabs by reverse transcription-polymerase chain reaction and SARS-CoV-2 specific IgG antibodies in serum by enzyme-linked immunosorbent assays and immunofluorescence tests. Discordant results were clarified by electrochemiluminescence immunoassays, a second enzyme-linked immunosorbent assay, or an in-house Luminex-based assay. This study included 4964 participants: 2482 children (median age, 6 [range, 1-10] years; 1265 boys [51.0%]) and 2482 parents (median age, 40 [range, 23-66] years; 615 men [24.8%]). Two participants (0.04%) tested positive for SARS-CoV-2 RNA. The estimated SARS-CoV-2 seroprevalence was low in parents (1.8% [95% CI, 1.2-2.4%]) and 3-fold lower in children (0.6% [95% CI, 0.3-1.0%]). Among 56 families with at least 1 child or parent with seropositivity, the combination of a parent with seropositivity and a corresponding child with seronegativity was 4.3 (95% CI, 1.19-15.52) times higher than the combination of a parent who was seronegative and a corresponding child with seropositivity. We observed virus-neutralizing activity for 66 of 70 IgG-positive serum samples (94.3%). In this cross-sectional study, the spread of SARS-CoV-2 infection during a period of lockdown in southwest Germany was particularly low in children aged 1 to 10 years. Accordingly, it is unlikely that children have boosted the pandemic. This SARS-CoV-2 prevalence study, which appears to be the largest focusing on children, is instructive for how ad hoc mass testing provides the basis for rational political decision-making in a pandemic.

Recombinant Baculovirus-Produced Grass Carp Reovirus Virus-Like Particles as Vaccine Candidate That Provides Protective Immunity against GCRV Genotype II Infection in Grass Carp.

Grass carp reovirus (GCRV) leads to severe hemorrhagic disease in grass carp (Ctenopharyngodon idella) and causes economic losses in grass carp aquaculture. Recent epidemiological investigations showed that GCRV genotype II is the dominant subtype in China. Therefore, it is very important to develop a novel vaccine for preventing diseases caused by GCRV genotype II. In this study, we employed a bac-to-bac expression system to generate GCRV-II-based virus-like particles (VLPs). Previous studies have shown that the structural proteins VP3, VP4, and VP38 encoded by the segments S3, S6, and S10 of type II GCRV are immunogenic. Hence, the GCRV-VLPs were produced by co-infection of sf9 cells with recombinant baculoviruses PFBH-VP3, PFBH-VP4, and PFBH-VP38. The expressions of VP3, VP4, and VP38 proteins in GCRV-VLPs were tested by IFA and Western blot analysis. By electron microscopic observations of ultrathin sections, purified VLPs showed that the expressed proteins are similar in shape to GCRV genotype II with a size range from 40 nm to 60 nm. The immunogenicity of GCRV-VLPs was evaluated by the injection immunization of grass carp. The analysis of serum-specific IgM antibody showed that grass carp immunized with GCRV-VLPs produced GCRV-specific antibodies. Furthermore, injection with GCRV-VLPs increased the expressions of immune-related genes (IgM, IFN, TLR3, TLR7) in the spleen and kidney. In addition, grass carp immunized with a GCRV-VLPs-based vaccine showed a relative percent survival rate (RPS) of 83.33% after challenge. The data in this study showed that GCRV-VLPs demonstrated an excellent immunogenicity and represent a promising approach for vaccine development against GCRV genotype II infection.

Enhanced immersion vaccination through hyperosmotic treatment in the largemouth bass (Micropterus salmoides)

This study aims to evaluate the effects of immersion vaccination through hyperosmotic treatment, and determine the optimum hyperosmotic salinity for largemouth bass (Micropterus salmoides) by determining immune responses and protective efficacies post vaccination. For hyperosmotic immersion (HI), M. salmoides were immersed in three hyperosmotic salinities (5, 10 and 20‰ salinity) respectively, then transferred into normal freshwater containing inactivated Aeromonas veronii. For direct immersion (DI) group, fish were directly immersed in freshwater with the same concentration of bacterin. Non-specific immune responses were evaluated based on the analysis of nonspecific humoral substances and immune related genes (interleukin 1β, CD40, IL-8 and tumor necrosis factor α), the results showed that serum lysozyme and myeloperoxidase activities were appreciably up-regulated earlier and maintained higher levels in HI groups than DI group, highest activities were observed in 10‰ salinity HI group. The expression dynamics of immune related genes had similar trend with nonspecific humoral substances. For specific cellular immune responses, the percentages of surface membrane immunoglobulin-positive (sIg+) cells in DI group did not show any significant increase until at 14 day post immunization (dpi) and peaked at 35 dpi (23.17%), sIg+ cells in HI groups showed a marked rise 7 days earlier than DI group, then steadily increased and peaked at 28 dpi (5‰ salinity: 25.83%; 10‰ salinity: 32.03%; 20‰ salinity: 24.17%). The serum specific antibody level showed obvious increase in all vaccination groups since 7 dpi, it reached the peak at 28 dpi in HI groups (5‰ salinity: OD = 0.49; 10‰ salinity:OD = 0.71; 20‰ salinity:OD = 0.50), one week earlier than DI group. Both sIg+ cells and specific antibody level were highest in the 10‰ salinity HI group. The relative percent survival (RPS) was 68.42%, 89.42% and 63.16% in 5, 10 and 20‰ salinity HI groups, respectively, higher than this in DI group (47.37%). These results demonstrated that HI at 10‰ salinity could efficiently enhance the immune responses of M. salmoides and provide higher RPS. This has significant value for hyperosmotic immersion with bacterin in M. salmoides.

Astragalus Polysaccharides Enhance the Immune Response to Newcastle Disease Virus Vaccination in Chickens

This study was conducted to evaluated the effects of Astragalus polysaccharide on immune responses of chickens immunized with NDV Lasota vaccine. One hundred chickens at one-day-old( Ross breed) were brought , on five day of age, the average maternal serum Abs was measured by hemaglutination inhibition (HI) test and the titer was 3.43 log, and then divided into five equal groups in each group twenty chicks and on day 10 of age the Astragalus polysaccharide high concentration (APSH) group were given 0.5 ml at 400mg /100ml , the Astragalus polysaccharide medium concentration (APSM )group were given 0.5 ml at 200mg /100ml, the Astragalus polysaccharide low concentration (APSL) group were given 0.5 ml at 100mg /100ml , Vaccine control (VC) group were given 0.5 ml normal saline only, Negative control (NC ) group were given normal saline only , all groups orally administrated for four days. At 14 day of age, all chickens with the exception of NC group , were vaccinated with ND Lasota by Intraocular and Intranasal methods. Four blood sample from all groups were aspirated from jugular vein at 0h (before vaccination),6h,12h and 24h(after vaccination ) for determine of Chicken IL-6(Interleukin 6) by RT-PCR. On day 20and 30 of age after vaccination , three chicken were sampled randomly from each group to detect specific serum Antibody titer of NDV by HI test. On days 1,7,14,21after vaccination, four serum Samples from the same of chicken to determines IgA level by sandwich ELISA test. The cellular and humeral response including the production of cytokine IL-6 ( by RT-PCR test) were measured on 0h(before vaccination),6h,12h and 24h( after vaccination )and IgA antibody ( by ELISA test ) were measured on 1d,7d,14d,and 21days after vaccination and serum antibody titer Specific to NDV( by HI test ) were measured at age 20, and 30 days after vaccination evaluated by series of experiments. Results revealed that all the polysaccharide groups were numerically increased in antibody levels, the expression of IL-6and IgA level ,but three parameter were significant (𝑃 &lt; 0.05) in the APSH group compared to corresponding (VC) vaccinated group and( NC) non –vaccinated group. These results suggest that orally administered APS could significantly enhance the efficacy of NDV vaccination and has important implications for the further use of APS as a novel adjuvant.

Astragalus Polysaccharides Enhance the Immune Response to Newcastle Disease Virus Vaccination in Chickens

This study was conducted to evaluated the effects of Astragalus polysaccharide on immune responses of chickens immunized with NDV Lasota vaccine. One hundred chickens at one-day-old( Ross breed) were brought , on five day of age, the average maternal serum Abs was measured by hemaglutination inhibition (HI) test and the titer was 3.43 log, and then divided into five equal groups in each group twenty chicks and on day 10 of age the Astragalus polysaccharide high concentration (APSH) group were given 0.5 ml at 400mg /100ml , the Astragalus polysaccharide medium concentration (APSM )group were given 0.5 ml at 200mg /100ml, the Astragalus polysaccharide low concentration (APSL) group were given 0.5 ml at 100mg /100ml , Vaccine control (VC) group were given 0.5 ml normal saline only, Negative control (NC ) group were given normal saline only , all groups orally administrated for four days. At 14 day of age, all chickens with the exception of NC group , were vaccinated with ND Lasota by Intraocular and Intranasal methods. Four blood sample from all groups were aspirated from jugular vein at 0h (before vaccination),6h,12h and 24h(after vaccination ) for determine of Chicken IL-6(Interleukin 6) by RT-PCR. On day 20and 30 of age after vaccination , three chicken were sampled randomly from each group to detect specific serum Antibody titer of NDV by HI test. On days 1,7,14,21after vaccination, four serum Samples from the same of chicken to determines IgA level by sandwich ELISA test. The cellular and humeral response including the production of cytokine IL-6 ( by RT-PCR test) were measured on 0h(before vaccination),6h,12h and 24h( after vaccination )and IgA antibody ( by ELISA test ) were measured on 1d,7d,14d,and 21days after vaccination and serum antibody titer Specific to NDV( by HI test ) were measured at age 20, and 30 days after vaccination evaluated by series of experiments. Results revealed that all the polysaccharide groups were numerically increased in antibody levels, the expression of IL-6and IgA level ,but three parameter were significant (𝑃 &lt; 0.05) in the APSH group compared to corresponding (VC) vaccinated group and( NC) non –vaccinated group. These results suggest that orally administered APS could significantly enhance the efficacy of NDV vaccination and has important implications for the further use of APS as a novel adjuvant.

Source investigation on a familiar cluster of coronavirus disease 2019 in Dandong city of Liaoning Province

To Track the source of the infection through an investigation of a clustering of coronavirus disease 2019(COVID-19), and provide scientific basis and Strategy for the effective control of the aggregated epidemic situation of COVID-19. Field epidemiological method was used to survey the cases and related close contacts in a family clustering epidemic of COVID-19 in Dandong city of Liaoning Province. We obtained survey data for a descriptive analysis.Real time RT-PCR technique was used to detect 2019-nCoV nucleic acid in samples collected from cases and related close contacts combined with serum specific antibody detection. A total of 3 confirmed cases and 2 asymptomatic infection cases were discovered in the clustering epidemic, with 34 close contacts.Of eight close family contacts visiting from other province, one patient was on the same flight as the confirmed case, and her antibody IgG was positive. The family clustering was caused by past infection case who visited her friend through Wuhan from other provinces to local area.

LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR DETECTION OF C. BURNETII IN BULGARIA

Q fever, which is caused by Coxiella burnetii, a small, pleomorphic intracellular bacterium, is the most widespread zoonosis in the world. The chronic form of the disease can lead to disability and death. Rapid diagnosis of Q fever is needed in order that effective treatment can be initiated. The conventional retrospective diagnosis of Q fever, based on serology, is useless for the treatment of afflicted patients. Thus, molecular methods have been created to close the diagnostic gap between the onset of the disease and the presence of specific antibodies in serum. A polymerase chain reaction is a suitable and reliable method with high sensitivity and specificity, but it requires expensive equipment and post-amplification protocol. Loop-mediated isothermal amplification (LAMP) is an isothermal technique, conducted at constant temperature that can amplify a negligible amount of DNA to more than 109 copies within one hour, using special primers and polymerase. We have tested the sensitivity and specificity of LAMP in the detection of C. burnetii. The mean positive rate of LAMP and polymerase chain reaction in patients was 100% and 74%, respectively. LAMP reacted negatively with non-C. burnetii pathogens and non-infected blood samples. We conclude that LAMP is a sensitive and specific technique for the detection of C. burnetii and has advantages over serological methods and PCR that make it attractive for diagnosing Q fever in countries around the world.

Clinical Characteristics, Serological and Radiological Longitudinal Changes of Patients with COVID-19.

Since December 2019, an outbreak of coronavirus disease 2019 (COVID-19) began in Wuhan and rapidly spread globally. The speed and scope of the spread of COVID-19 makes it urgent to define clinical characteristics, serological and radiological changes of the affected patients. Seven patients with laboratory-confirmed COVID-19 who were admitted to the Third Affiliated Hospital of Sun Yat-Sen University Yuedong Hospital from January 2020 to March 2020 were retrospectively enrolled and their clinical features, serological and radiological longitudinal changes were analyzed. Among the 7 patients, all (100%) had a clear epidemiological history. The most common symptoms were respiratory symptoms 6 (85.7%), and only 2 (28.6%) of the patients had fever at their first visit. The cohort included 4 (57.1%) common types and 3 (42.9%) severe types. Two (28.6%) common type patients developed to severe type in a short time. All of the 7 patients (100%) had abnormal liver function, normal renal function, and normal procalcitonin. The detection time of specific antibody in 7 patients was 5 - 13 days after symptoms. Before the specific antibody could be detected, the absolute value of lymphocytes decreased in 2 (28.6%) common type cases transferred to severe type cases accompanied with obvious progress in pulmonary imaging. The phenomenon of decreased albumin and elevated globulin occurred in 6 patients (85.7%). The predominant pattern of lung lesions observed was bilateral (71.4%) and mainly near the pleura at the first diagnosis. Bilateral pulmonary involvement occurred in 6 cases (85.7%) during the course of disease. In 4 cases (57.1%) with obvious pulmonary lesions, the absolute value of lymphocytes decreased, albumin decreased, and globulin increased during the course of the disease. Serum specific antibodies can be detected within 2 weeks of onset. Close observation of the dynamic changes of absolute value of blood lymphocytes, serum albumin, and globulin which were related to pulmonary imaging changes in patients will contribute to assessment of COVID-19.

Пищевая и ингаляционная сенсибилизация при атопическом дерматите

Aim: to study the association of food and inhalation sensitization with the severity of atopic dermatitis (AtD) in order to improve its treatment and prevention methods. Patients and Methods: the study included 50 patients with AtD from 2 to 44 years old. The study included clinical and laboratory research methods. The survey analyzed the history and anamnesis vitae, estimated the age at the AtD onset, determined the nature of concomitant pathology, the possible causes of cutaneous exacerbation, the allergic severity in the family history, the severity score on the SCORAD (Scoring of Atopic Dermatitis) scale and the efficacy of previous therapy. The immunological examination consisted of the quantitative determination of specific IgE and IgG antibodies in the blood serum to inhaled and food allergens by immunochemiluminescence assay on an IMMULITE 2000 XPi analyzer. Results: according to the detection frequency of IgE to food allergens in patients with AtD: the leading positions were taken by the chicken egg (egg white in 26% of cases, egg yolk in 20%); IgE to banana was detected in 17% of cases; in 12% — to cow’s milk, more common — to the β-lactoglobulin fraction. When studying the activity of inhaled allergens, respiratory sensitization to the cat epithelium was detected in 26% of cases, and to black birch — in 24%. Hypersensitivity to herbs (wormwood, herbal mixture) and house dust allergens (8%, respectively) was found much less common. In severe AtD, IgE to respiratory allergens prevailed: at n=38, the IgE concentration to cat epithelium and black birch was 12.4±1.4 IU/mL each. Besides, there was a significant sensitization to food allergens, namely, the IgE concentration to egg white — 5.4±0.9 IU/mL. Conclusion: the study revealed the sensitization of patients with AtD to food (mainly egg white, egg yolk, banana and cow’s milk) and respiratory (mainly cat epithelium, black birch) allergens. Determination of the IgE concentration in the blood serum of patients showed that there was a significant concentration increase to egg white, cat and black birch in severe AtD. Therefore, it is advisable to test for specific allergens in all patients with moderate to severe AtD. KEYWORDS: food sensitization, inhalation sensitization, atopic dermatitis, allergy, allergens, antibodies, immunochemiluminescence assay, IgE, triggers. FOR CITATION: Vasilyeva E.A., Rusak Yu.E., Efanova E.N., Lebedeva L.N. Food and inhalation sensitization in atopic dermatitis. Russian Medical Inquiry. 2021;5(1):17–20. DOI: 10.32364/2587-6821-2021-5-1-17-20.

Serum of Mice Immunized with Mt1-MMP Metalloproteinase Reduces Migration Potential of Pancreatic Cancer Cells

Expression levels of matrix metalloproteinases, in particular MT1-MMP, are elevated in pancreatic cancer (PC) cells, and this is associated with increased tumor proliferation, invasion, and migration. MT1-MMP is considered a promising target for drug therapy of PC, but the use of inhibitors and therapeutic antibodies to MT1-MMP is limited because maximal efficiency is only observed in a narrow time interval, at the early asymptomatic stages of the disease. This problem could be solved by immunization to MPs at the moment of detection of the primary tumor. This therapeutic effect could be provided by specific antibodies that can be re-produced in case of relapses. Here, we selected the optimal mode for immunization of mice with MT1-MMP fragments that allows us to obtain a high titer of specific antibodies in the blood serum. The obtained antiserums effectively inhibited MT1-MMP enzymatic activity, migration of PANC-02 PC cells through the collagen matrix, and activation of the main inducers of epithelial -mesenchymal transition, TGF-β and MMP-2. These results maybe useful in the development of drugs for PC treatment, and the approach we propose might form the basis for design of antitumor drugs with prolonged action.